Hossein Asgarian; Mahdi Shabani; Parvaneh Vosoogh; Ramazan Ali Sharifian; Soheila Gharagozlou; Jalal Khoshnoodi; Tahereh Shahrestani; Mahin Kordmahin; Abdolfattah Sarrafnejad; Mahmood Jeddi Tehrani; Hodjatallah Rabbani; Fazel Shokri
Volume 2, Issue 4 , December 2005, , Pages 182-190
Abstract
Background: The Wilm’s tumor gene 1 (WT1) encodes a zinc finger transcription factor that is inactivated in a subset of Wilm’s tumors. It plays a crucial role in growth, proliferation and development of some embryonic and adult organs. WT1 is expressed as a tumor associated antigen (TAA) ...
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Background: The Wilm’s tumor gene 1 (WT1) encodes a zinc finger transcription factor that is inactivated in a subset of Wilm’s tumors. It plays a crucial role in growth, proliferation and development of some embryonic and adult organs. WT1 is expressed as a tumor associated antigen (TAA) in various types of solid and hematopoietic malignancies and can be employed as a useful marker for targeted immunotherapy and monitoring of minimal residual disease (MRD). Objective: To investigate the profile of WT1 gene expression in Iranian patients with acute myeloblastic leukemia. Methods: RT-PCR method was used to determine the WT1 gene expression in bone marrow (BM) and/or peripheral blood (PB) samples from 11 patients with AML and PB samples of 36 normal subjects. Isolated cells from all patients were immunophenotyped by flow cytometry. Results: The leukemic cells from 10 patients (91%) were found moderately or strongly positive for WT1 expression whereas only 3 out of 36 normal subjects expressed WT1 at very low levels. A highly significant correlation was observed for WT1 expression between paired BM and PB samples of the AML patients. Conclusion: Our results indicate that WT1 is expressed in the majority of Iranian AML patients and may be employed for screening and monitoring of minimal residual disease in these patients.
Fatemeh Hajighasemi; Soheila Gharagozlou; Nasrin Moheghi; Roya Ghods; Jalal Khoshnoodi; Fazel Shokri
Volume 2, Issue 3 , September 2005, , Pages 125-133
Abstract
Background: There are two subclasses of human IgA (IgA1 and IgA2) that differ in antigenic properties and in chemical composition. The constant domains of α1 and α2 heavy chains have >95% sequence homology though major structural differences exist in the hinge region. Quantitation of IgA ...
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Background: There are two subclasses of human IgA (IgA1 and IgA2) that differ in antigenic properties and in chemical composition. The constant domains of α1 and α2 heavy chains have >95% sequence homology though major structural differences exist in the hinge region. Quantitation of IgA subclass levels depends on the availability of monoclonal antibodies (MAbs) specific for conserved conformational or linear epitopes restricted to each subclass. Objective: To produce, select and characterize monoclonal antibodies specific for human IgA2. Methods: Splenocytes from BALB/C mice immunized with a human IgA2 myeloma protein were fused with SP2/0 myeloma cells. Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by limiting dilution assay. Antibody (Ab) secreting cells were screened by enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAbs was further analyzed, using a panel of purified myeloma proteins and some animal sera by ELISA and immunoblotting. The affinity constant (Kaff) was also determined by ELISA. Results: Four murine hybridoma clones designated 2F20G5, 2F20B5, 3F20E3 and 6F20H11 were obtained that secreted MAbs specific for the human IgA2. 2F20G5 and 6F20H11 MAbs react with linear epitope(s) while 2F20B5 and 3F20E3 react with conformational epitope(s) located to human IgA2 subclass. 2F20G5 MAb displays a weak cross-reactivity with monkey and rabbit sera and a strong cross-reactivity with cat and dog sera while the other three MAbs showed no cross-reactivity with the animal sera tested. Conclusion: These MAbs, especially 6F20H11 with high affinity constant (6.03 ×109 M-1) are useful tools for quantitation of human IgA2 subclass levels in various diseases. Cross-reactivity of 2F20G5 MAb with some animalsera suggests phylogenic conservation ofthe epitope recognized by this MAb.